• Analytical Science and Technology
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• v.31 no.5
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• pp.208-218
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• 2018

Autophagy is a cellular process whereby cytosolic materials or organelles are taken up in a double-membrane vesicle structure known as an autophagosome and transported into a lysosome for degradation. Although autophagy has been studied at the genetic, cellular, or biochemical level, systematic ultrastructural quantitative analysis of autophagosomes during the autophagy process by using transmission electron microscopy (TEM) has not yet been reported. In this study, we performed ultrastructural analysis of autophagosomes in wild-type (WT) mouse embryonic fibroblasts (MEFs) and autophagy essential gene (atg5) knockout (KO) MEFs. First, we performed ultrastructural analysis of autophagosomes in WT MEFs compared to atg5 KO MEFs in basal autophagy or starvation-induced autophagy. Although we observed phagopore, early, late autophagosomes, or autolysosomes in WT MEFs, atg5 KO MEFs had immature autophagosomes that showed incomplete closure. Upon starvation, late autophagosomes accumulated in WT MEFs while the number of immature autophagosomes significantly increased in atg5 KO MEF indicating that atg5 plays an important role in the maturation of autophagosomes. Next, we examined autophagosomes in the cell model expressing polyQ-expanded N-terminal fragment of huntingtin. Our TEM analysis indicates that the number of late autophagosomes was significantly increased in the cells expressing the mutant huntingtin, indicating that improving the fusion of autophagosome with lysosome may be effective to enhance autophagy for the treatment of Huntington's disease. Taken together, the results of our study indicate that ultrastructural and quantitative analysis of autophagosomes using TEM can be applied to various human cellular disease models, and that they will provide an important insight for cellular pathogenesis of human diseases associated with autophagy.

• Applied Microscopy
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• v.31 no.2
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• pp.157-165
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• 2001

Sertoli cells in the normal adult testis are nondividing cells, which are relatively inconspicuous on cross section of the seminiferous tubule and comprise about 10% to 15% of the tubular cellular elements. Ultrastructurally, Sertoli cells have characteristic nucleoli, plasma membrane, and cytoplasmic components. The plasma membrane has two types of intercellular junctions which are developed at puberty: junctions between adjacent Sertoli cells and Sertoli cell-germ ceil junction. However, the ultrastructural findings of Sertoli cells in human fetus is not fully elucidate yet. In the present study, human fetal testes ($14\sim27$ weeks) obtained from artificially induced abortions legally without gross malformation were studied using transmission electron microscopy to make clear the differentiation process of Sertoli cells in human. In human fetal testes from 14 weeks to 27 weeks, the cell junctions of Sertoli-germ cells and Sertoli-Sertoli cells are desmosome like structure and not tight junction or desmosome. The Overall intracytoplasmic organelles of Sertoli cells are relatively sparse. The mitochondrias are relatively abundant but no developed cristae. And the rough endoplasmic reticuli are abundant and smooth endoplasmic reticuli are sparse. The amount of lipid droplets are regularly observed in human fetal Sertoli cells. No microfilaments or Charcot-Bottcher's crystalloids are present. From the results, Sertoli cells in human fetal testes are somewhat different ultrastructural findings with puberty or adult. However, to make clear the differentiation process of Sertoli cells in human, further study for 28 weeks to puberty is required.

• The Journal of Korean Medicine
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• v.18 no.1
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• pp.430-445
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• 1997

In order to know the effects of Calculus $Bovis{\cdot}F디$ Ursi aqua-acupuncture ($C{\cdot}F$ aqua-acupuncture) to the liver damage, we carried out the treatment of $C{\cdot}F$ aqua-acupuncture and injection of Kam-Du-Tang on the liver damaged rat which is induced by aconitine, and then ultrastructural study has been to the liver tissue. The results were as follows: 1. In the hepatic cell of normal group, the nuclear envelope is round and electro-density of nucleoplasm is relatively low, and have one or two large nucleolus. Cell organelle is developed rough endoplasmic reticulum(RER) and many mitochondria, glycogen granules, smooth endoplasmic reticulum(SER) and Golgi apparatus. 2. In 24 hours control group, nuclear envelope is very irregular, RER and most of cell organell is shown that destroyed by aconitine. This is similar to the 48 hours control group. In 72 hours control group, RER is recovered somewhat but cristae of mitochondria is not seen the shape 3.In 24 hours $C{\cdot}F$ aqua-acupuncture group, nuclear envelope is very irregular and cristernae is very dilatable and many mitochondria is dilationed and the cristae is not definite like the control group. In 48 hours $C{\cdot}F$ aqua-acupuncture group, nuclear envelope is relatively round and chromatin is regular. Cristernae of RER and ribosme is somewhat imperfect but is similar the normal group, and the cristae of mitchondria is shown definitely like the normal group. In 72 hours $C{\cdot}F$ aqua-acupuncture group, nuclear envelope is round and cell organell is similar to normal group. 4. In 24 hours Kam-Du-Tang group, nuclear envlope is very irregular and cell organelles are destroyed by the effects of aconitine. In 48 hours Kam-Du-Tang group, nuclear envelope is somewhat irregular but chromatin is relatively regular. RER is seperated regularly through the cytoplasm, but cannot make the structure of lamina, and cistennae is very dilationed. many of glycogen granules is seperated regularly. In 72 hours Kam-Du-Tang group, RER makes the lamina. Mitochondria is a little but inner space is dilationed so the shape of is not definite and a lot of glycogen granules are accumulated in several places. It see the above result, hepatic cell that is damaged by aconitine is effected chiefly by the $C{\cdot}F$ acupuncture and effect of acupuncture is similar to the detoxicated effect of Kam-Du-Tang.

• Applied Microscopy
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• v.8 no.1
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• pp.17-22
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• 1978

Two cases of Histiocytosis X, a Hand-Schuller-Christian disease and a Letterer-Siwe disease, are,examined light and electron microscopically. Many of the histiocytes without appreciable lipid contained numerous rod-shaped structure identical to Langerhan's granules in the cytoplasm. The significance of this structure was discussed together with review of literature.

• Mycobiology
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• v.38 no.4
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• pp.328-330
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• 2010

In this study, we demonstrate that equol has fungicidal activities against Candida albicans. The minimum inhibitory and minimum fungicidal concentrations of equol against C. albicans were 516 and $1,032{\mu}M$, respectively. Two separate viability assays found that equol changed the integrity of the C. albicans cell membrane, possibly by formation of membrane lesions. Scanning electron microscopy demonstrated ultrastructural changes.

• Proceedings of The Korean Cleft Lip And Palate Association
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• 2003.06a
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• pp.38-38
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• 2003

• Proceedings of the Korean Society of Toxicology Conference
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• 2002.11b
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• pp.148-148
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• 2002

This study is to characterize the microscopical and ultrastructural changes in chronic exposure of Microcystin-LR (MCLR), a cyclic heptapeptide hepatotoxin, comparing to those in acute lethal toxicity. Female ICR mice were injected intraperitoneally with 10, 20, 30,$\mu\textrm{g}$/kg of MCLR every 3 day for 27 days.(omitted)

• Applied Microscopy
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• v.25 no.4
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• pp.26-51
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• 1995

The present study was designed to examine effect of long term weight-training on aging atrophy in the rat skeletal muscle. Male rats of 8, 15, and 24 month old were used. Each age groups included control and weight-training for 5 months by using body press apparatus. The histo- and cytochemical, ultrastructural and stereological changes in aging skeletal muscles of the rat were observed in the present study. During the training period the body weight and muscular weight in all groups except the rectus femoris and the gastrocnemius in young age groups remained constant, but muscular weights were increased in the rectus femoris and the gastrocnemius muscles in young age groups. In trained rat, the volume density of muscle fiber type IIA and IIB were increased, but those of type IIC was decreased. Type I remained constant in 8 and 15 month old age groups, but reduced in the tibialis anterior and the gastrocnemius muscles in the 24 month old groups. Some histotological and ultrastructural changes associated with age were found: numerical increase of cytiplasmic vacuoles, lysosomes, lipofuscins, and irregularity of myofibrils. At 24 month old groups some unusual formation of contraction band and muscle splitting were observed. After weight-training, ultrastructural degenerative changes occured in the type I muscle fiber, such as splitting of muscle fiber, disorganization of myofilaments, swelling of mitochondria, accumulation of many lipid droplets, appearance of many lysosomes and residual bodies and necrotic fibers, in the old age groups. But, in the type II muscle fibers hypertrophy of muscle fiber appeared without any noticible damage as the type I. The activities of $Mg^{++}$ -ATPase decreased with age and this enzyme activities in the trained rat were significantly decreased with age. Activities of the acid phosphatase were increased with age and significantly in the trained rat. In stereological analysis, volume density of the myofibrils and the tubular system were increased, on the other hand there mitochondrial capacity was decreased. These experimental results suggested that old rats are not susceptible to be affected by weight-training as young rats, and that physical capacity of the rats must be considered when old rats are exercised for training.

• Applied Microscopy
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• v.27 no.3
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• pp.265-279
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• 1997

This study aims to demonstrate the effect of chitosan, one of the natural chelator, on the ultrastructural changes in the mouse liver caused by cadmium. A total of 60 healthy ICR that weighted 30 gm $({\pm}2gm)$ was used for experiment. The experimental group was divided into three groups; group A, B, and C. The group A and B administrated cadmium (4.0 mg/kg) to the intraperitoneal after pretreated with chitosan (0.5% solution) for 30 and 7 days, respectively. Each group was observed at 12, 24, 48, 72 hours and one week after injected cadmium. The results were as follows: 1. Group A The nuclear membrane and the chromatin were normal shapes at overall the time. The inner and outer membranes of the mitochondria damaged a little but almost normal in shapes. And electron-density showed slightly compacted. some enlarged rER (rough endoplasmic reticulum) showed at 12 hours. At 48 hours, typical lamellae of the rER were reformed, and a lot of transvesicles observed around them. To 48 hours, sER (smooth endoplasmic reticulum) was slightly dilated. From 72 hours, sER rehalizated in normal shape. 2. Group B Nuclear membranes were rounded-shape and chromatin showed evenly. To 72 hours, a lot of mitochondria observed around rER and development of cristae showed weakly. But at one week, cristae were clear and electron-density of matrix showed high. At 72 hours lamellae of rER showed some broken, but were reformed at one week. Also at one week, glycogen granules evenly showed over cytoplasm. 3. Group C At 12 hours, Nucleus showed the condensation of nuclear membrane and clear condensation at 24 hours. However, nuclear membrane had a slightly rounded-shape from 72 hours. From 12 hours to the one week, mitochondria showed the dilation of inner cavity and weak development of cristae. Also electron-density of matrix was a little low. Occasionally, destruction of inner and outer membrane observed at one week. The dilation of cisternae and destruction of lamellae of rER showed from 12 to 48 hours. From 72 hours, rER showed slightly dilated only. And lamella observed at one week. In sER, dilation of inner cavity was observed during whole period. These results suggest that chitosan attenuates the toxic effect of the cadmium in the mouse liver.

• Journal of Chest Surgery
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• v.33 no.5
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• pp.377-384
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• 2000

Background: Numerous studies of safe, long term preservation for lung transplantation have been performed using ex vivo models or in vivo single lung transplantation models. However, a safe preservation time which is applicable for clinical use is difficult to determine. We prepared LPDG solution for lung preservation study. In this study we examined the efficacy of LPDG(low potassium dextran glucose) solution in 24-hour lung preservation by using a sequential bilateral canine lung allotransplant model. Material and Method: Seven bilateral lung transplant procedures were performed using weight-matched pairs(24 to 25kg) of adult mongrel dogs. The donor lungs were flushed with LPDG solution and maintained hyperinflated with 100% oxygen at 1$0^{\circ}C$ for a planned ischemic time of 24 hours for the lung implanted first. After sequential bilateral lung transplantation, dogs were maintained on ventilators for 3 hours: arterial resistance were determined if the recipients hourly after bilateral reperfusion and compared with pretransplant-recipient values, which were used as controls. After 2hours of reperfusion, the chest X-ray, computed tomogram and lung perfusion scan were performed for assessmint of early graft lung function. Pathological examinations for ultrastructural findings of alveolar structure and endothelial structure of pulmonary artery were performed. Result: Five of seven experiments successfully finished the whole assessments after bilateral reperfusion for three hours. Arterial oxygen tension in the recipients was markedly decrased in immediate reperfusion period but gradually recovered after reperfusion for three hours. The pulmonary artery and pulmonary vascular resistance showed singificant elevation(p<0.05 versus control values) but also recovered after reperfusion for three hours(p<0.05 versus immediate period value). The ultrastructural findings of alveolar structure and endothelial structure of pulmonary artery showed reversible mild injury in 24 hours of lung perservation and reperfusion. Conclusion : This study suggests that LPDG solution provides excellent preservation in a canine model in which the dog is completely dependent on the function of the transplanted lung.